Discussed about Sources of Heavy metals , Sources of Heavy metals , Bioremediation, Biosorption by Fungi, Algae, Bacteria , Factors affecting Biosorption , Heavy metals relation with human beings

1. BIOSORPTION OF
HEAVY METALS

2. CONTENTS
1.Introduction ……………………………………………………………………….…1
1.1 Types of Heavy Metals with Atomic Number ……………………….…1-2
2.Sources of heavy metals………………………………………………………………2
2.1 Anthropogenic Source……………………………………………………....2
2.2 Natural Source…………………………………………………………….…2
2.3 Effect of heavy metals ……………………………………………………….3
2.4 Toxic range of Heavy Metal……………………………………………….3-4
3.Bioremediation…………………………………………………………………………4
3.1 Procedures for removing heavy meatal…………………………………..4-6
4.Biosorption ……………………………….…………………………………………...6-7
4.1 Biosorption Mechanism…………………………………………………….7-9
5. Microorganisms in metal absorption ……………………………………………..…10
5.1 Biosorption by Fungi…………………………….………………………..…10
5.2 Biosorption by Algae and Moss…………………….………………...…10-11
5.3 Biosorption by Bacteria …………………………………….…………..11-12
6.The factors affect the Biosorption Process……………………………….………12-13
6.1 Microorganisms used in Biosorption process……………………….….13-14
7.Heavy Metals – Humans …………………………………………………………...14-15
8.Conclusion……………………………………………………………………………….16
9.References ……………………………………………………………………………….17

3. ABSTRACT
Industrialization has led to introduction of heavy metals in the environment. Heavy metals are
known to persist in the environment and become a risk for organisms. Micro-organisms are present
in industrial effluents. They have adopted different strategies to cope up with harmful effects of
these metals. The strategies can be metabolism dependent or independent. Once such strategy is
biosorption which is binding of metal ions with metal binding proteins present on the cell wall.
Biosorption is exhibited by bacteria, algae, fungi and yeasts. Not only living organisms but also
residuals of dead bodies of microorganisms absorbs heavy metals. Different factors affect the rate
of biosorption which include temperature, pH, nature of biosorbents, concentration of biomass,
initial metal ion concentration and metal affinity to biosorbents,
Keywords: Microorganisms, Heavy Metals, Bioremediation, Biosorption, Industrial effluent

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1.INTRODUCTION
Heavy metals are defined as those elements having an atomic number greater than 20 and atomic
density above 5g cm-3 and must exhibit the properties of metal. The Heavy metals can be broadly
classified into two categories essential and nonessential heavy metals. Essential heavy metals are
those required by living organisms for carrying out the fundamental process like growth,
metabolism, and development of different organs. There are numerous essential heavy metals like
Cu, Fe, Mn, Co, Zn, and Ni required by plants as they form cofactors that are structurally and
functionally vital for enzymes and other proteins. Essential elements are often required in trace
amounts in the level of 10-15ppm and are known as micronutrients. Nonessential heavy metals
like Cd, Pb, Hg, Cr, and Al are not required by plants even in race amounts for any of the metabolic
processes.
Heavy metals are a group of metals and metalloid that have relatively high density and are toxic
even at ppm levels. Examples include Pb, As, Hg, Cd, Zn, Ag, Cu, Fe, Cr, Ni, Pd, and Pt. These
metals are released into the environment by both natural and anthropogenic sources such as
industrial discharge, automobiles exhaust, and mining. Unlike organic pollutants heavy metals are
nonbiodegradable and have tendency to accumulate in living beings. In fact, most of them are
known due to long term and continuous exposure to heavy metals. Since they are non-degradable
and tend to bioaccumulate, suitable methods need to be established for their efficient removal from
the environment.
1.1Types of Heavy metals with Atomic Number
Table -1
Heavy Metal Atomic No
Antimony 51
Arsenic 33
Barium 56

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Cadmium 48
Copper 29
Chromium 24
Lead 82
Manganese 25
Mercury 80
Molybdenum 42
Nickle 28
Selenium 34
Uranium 92
2. SOURCES OF HEAVY METALS
The two main sources of Heavy metals are Anthropogenic and Natural Sources
2.1 Anthropogenic Source
Anthropogenic contaminations are pollution produced directly done by human beings like usage
of pesticides, wood preservatives, biosolids, ore mining, smelting, paints pigments, steel
industries, fly ash, fertilizers, kitchen appliances, automobile batteries etc.,
2.2 Natural Source
Naturally occurring pollutants also lead to source of heavy metals like weathering of minerals,
erosion and volcanic activates, forest fires and biogenic sources, particles released by vegetation.

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2.3 Effect of Heavy Metals
 On Soil and Plants
Plants growing on soil polluted with heavy metals show less seed germination, decreased
enzyme activity and in turn stunted growth, chlorosis, damage to root tips, reduced water
and nutrient uptake and impairment to enzyme functions due to Presence of heavy metal
in soil.
 On water bodies
Presence of lead in aquatic ecosystem decrease dissolved oxygen contents of water making
young fish more vulnerable to that change. The deficiency may cause blackening of tail of
young fish
 On Humans
Effect may be acute to chronic causing gastrointestinal disorder stomatitis, diarrhea,
hemoglobinuria, vomiting, convulsion
2.4 Toxic range of Heavy Metal
 Cadmium (Cd)
is highly toxic even at very low concentration a long term exposure to it may cause renal
dysfunction, lung disease, osteomalacia, Osteoporosis and myocadiac dysfunction, severe
exposure may lead to pulmonary edema and even death.
 Lead (Pb)
Accumulated in soft tissue of children causes stunted growth, damage to nervous system,
learning disabilities and anti-social behavior. Lead (Pb) accumulation may also lead to
decrease in hemoglobin production, cardio vascular disorder and injury to central and
peripheral nervous system.
 Copper (Cu)
Is an essential element but its higher concentration in drinking water may cause liver
cirrhosis anemia and kidney damage.

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 Zinc (Zn)
Is a component of many enzymes like carbonic anhydrase alkaline phosphate, alcohol
dehydrogenase etc., high concentration of Zn may result in growth and reproduction
impairment Clinical signs of Zn toxicity is diarrhea, vomiting Anemia eye irritation and
bloody urine
 Mercury (Hg)
Is highly known toxic metal for human toxicity symptoms of Hg depend upon its form of
ingestion. Organic form – Spontaneous abortion and gastrointestinal disorders
 Arsenic (As)
High levels of Arsenic may cause death. Arsenic coagulates with proteins, forms
complexes with some enzymes and inhibits ATP production.
3.BIOREMEDIATION
Bioremediation is a waste management technique that involves the use of organism to remove or
neutralize pollutants from a contaminated site according to Environmental Protection Agency
Bioremediation is a “treatment that uses naturally occurring organisms to break down hazardous
substances into less toxic or non-toxic substances”. However, if the biological activity needed to
degrade a particular contaminant is not present at the site, suitable microbes from other locations,
called exogenous microorganisms can be introduced and nurtured. Other technologies being
demonstrated are phytoremediation or the use of plants to clean up contaminated soils and ground
water, and fungal remediation, employs white-rot fungus to degrade contaminants.
*Biologically- based remediation detoxifies hazardous substances instead of merely transferring
contaminants from one environmental medium to another
*Bioremediation is generally less disruptive to the environment than excavation- based processes
The bioremediation technology most suitable for a specific site is determined by several factors,
such as site conditions indigenous microorganism population and the type, quantity and toxicity

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of contaminant chemicals present. Some treatment technologies involve the addition of nutrients
to stimulate or accelerate the activity of indigenous microbes. Optimizing environmental
conditions enhance the growth of microorganism and increase microbial population resulting in
improved degradation of hazardous substances.
Bioremediation is broadly discussed in two sectors in situ and Ex situ
*In situ-
at the site treatment of contaminated material in place
E.g.: - removal of lead, arsenic other heavy metals through phytoremediation, microbial
remediation
*Ex situ-
Away from site
Techniques involve physical removal of the contaminated material for treatment process
E.g.: -Bio-piles, soil treatment unit, compost pile, windrows
3.1 Procedures for removing heavy meatal
The commonly used procedures for removing metal ions from this environment both aquatic
streams and soil lands include Ion exchange, Chemical precipitation, Phytoremediation, Microbial
remediation.
 Chemical precipitation-
Precipitation of metals is achieved by the addition of coagulants such as alum, lime, iron
salts and other organic polymers. The large amount of sludge containing toxic compounds
produced during the process is the main disadvantages.

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 Phytoremediation-
Phytoremediation is the use of certain plants to clean up soil, sediment and water
contaminated with metals. The disadvantages include that it takes a long time for removal
of metals and the regeneration of the plant for further biosorption is difficult.
Phytoremediation can be used for removal of heavy metals and radio nuclear as well as for
organic pollutants. It is a novel cost effective, environment – and eco-friendly, in situ
applicable and solar driven remediation strategy. Plant generally handles the contaminants
without affecting topsoil, thus conserving its utility and fertility. The techniques included
in phytoremediation phytoextraction, Phytofiltration, phytodegradation, Rhizodegradation,
Phytovolatilization.
Even though phytoremediation is suggested for removal of heavy metals but still it have
some disbenefits in following these methods as –
*Phytoremediation is limited to the surface area and depth occupied by the roots.
*Slow growth and low biomass require a long-term commitment
*With plant-based systems of remediation it is not possible to completely prevent the
leaching of contaminants into the groundwater. The survival of the plants is affected by
the toxicity of the contaminated land and the general condition of the soil.
4. BIOSORPTION
Biosorption is a process that utilizes inexpensive live or dead biomass to sequester toxic heavy
metals. Bio absorbents are prepared from naturally abundant or waste biomass of algae, fungi,
moss or bacteria that have been killed. Conventional methods such as precipitation,
oxidation/reduction, ion exchange, filtration, membranes and evaporation are extremely expensive
or inefficient for metal removal. In this context the biosorption process has been recently
evaluated. Although biosorption is promising its mechanism is not well elucidated. Microbial
cells can accumulate heavy metals, radionuclides and organometalloid compound by a variety of

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physico- chemical and biological processes. Metabolism-independent binding or absorption to
living or dead cells, extracellular polysaccharides, capsules and slime layer to bacterial cells walls
and envelopes, cell walls of algae, fungi and yeast are efficient metal bio sorbents of heavy meals.
Sorption and complexation of dissolved metals based on the chemical activity of microbial
biomass are the base of the new biosorption technology for metal removal and recovery. Because
of their wide diversity of habitats, some bacterial have always been exposed to high concentrations
of metal ores. It is not surprising therefore that specific resistance mechanisms have evolved to
cope with toxic heavy metals. In addition to exposure from naturally occurring ores and during
exposure to high levels of metals in some environment.
Biosorption is possible by both living and nonliving biomass. Bioaccumulation is growth
dependent process and biosorption involves mechanism like ion exchange, chelation and
complexation and inorganic precipitation. The process may occur by hydrolysis and inorganic
deposition via adsorption by physical forces and ion entrapment in inter and intra-fibrillar
capillaries and spaces of the structural polysaccharide network as a result of diffusion through cell
walls and membranes. Several active groups of cell constituents like acetamido group of chitins,
structural polysaccharides of fungi amine, sulphahydral and carboxyl group in protein
phosphodiester, phosphate, hydroxyl in polysaccharides participate in biosorption.
4.1 Biosorption Mechanism
The complex structure of microorganism implies that there are many ways for the metal to be up
taken by the microbial cell. The biosorption mechanisms are various and are not fully understood.
They can be classified according to various criteria.
Transport across cell membrane
Heavy metal transport across microbial cell membranes may be mediated by the same mechanism
used to convey metabolically important ions such as potassium, magnesium and sodium. The
metal transport systems may become confused by the presence of heavy metal ions of the same
charge and ionic radius associated with essential ions. This kind of mechanism is not associated

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with metabolic activity. Basically, biosorption by living organisms comprises of two steps. First
a metabolism independent binding where the metals are bound to the cell walls and second,
metabolism dependent intracellular uptake, whereby metal ions are transported across the cell
membrane.
 Physical adsorption-
In this category, physical adsorption takes place with the help of Vander Walls forces
according to few hypotheses that uranium, cadmium, zinc, copper and cobalt biosorption
by dead biomasses of algae, fungi, and yeasts takes place through electrostatic interactions
between the metal ions in solutions and cell walls of microbial cells. Electrostatic
interactions have been demonstrated to be responsible for copper biosorption by bacterium
Zoogloeal ramigera and alga chlorella vulgaris for chromium bio sorption by fungi
Ganoderma lucidium and Aspergillus niger .
 Ion Exchange-
Cell walls of microorganisms contain polysaccharides and bivalent metal ions exchange
with the counter ions of the polysaccharides. For example, the alginates of marine algae
occur as salts of K+, Na+, Ca2+, and Mg2+. These ions can exchange with counter ions such
as Co2+, Cu2+, Cd2+ and Zn2+ resulting in the biosorptive uptake of heavy meals. The
biosorption of copper by fungi Ganoderma lucidium and Aspergillus niger was also uptake
by the ion exchange mechanism.
 Complexation-
The metal removal from solution may also take place by complex formation on the cell
surface after the interaction between the metal and the active groups. According to
hypothesis that biosorption of copper by Chlorella Vulgaris and Zoogloea ramigera take
place through both adsorption and formation of coordination bonds between metals and
amino and carboxyl groups of cell wall polysaccharides. Complexation was fund to be the
only mechanism responsible for calcium, magnesium, Cadmium, Zinc, Copper and
mercury accumulation by Pseudomonas syringae.

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 Precipitation-
Precipitation may be either dependent on the cellular metabolism or independent of it. In
the former case the metal removal from solution is often associated with active defense
system of the microorganisms. They react in the presence of a toxic metal producing
compounds. Which favor the precipitation process. In the case of precipitation not
dependent on the cellular metabolism it may be a consequence of the chemical interaction
between the metal and the cell surface.
Fig:1 Mechanisms of heavy metal uptake by microorganism
(Source: kanamarlapudi et al.,2018)
 Use of Recombinant bacteria for metal removal –
Metal removal by adsorbents from water and waste water is strongly influenced by
physico-chemical parameters such as ionic strength, pH and the concentration of
competing organic and inorganic compounds. Recombinant bacteria are being investigated
for removing specific metals from contaminated water. For example, a genetically
engineered Escherichia coli which express Hg2+ transport system and metallothionine (a
metal binding protein) was able to selectively accumulated 8 µmole Hg2+/g cell dry weight.
The presence of chelating agents Na+, Mg2+, and Ca2+ did not affect bioaccumulation.

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5.MICROORGANIMS IN METAL ABSORPTION
5.1 Biosorption by Fungi
Among microorganism fungal biomass offers the advantage of having a high percentage
of cell wall material that shows excellent metal binding properties. Many fungi and yest
have shown and excellent potential of metal biosorption particularly the genera Rhizopus,
Aspergillus, Streptoverticillum, and Saccharomyces.
Polysaccharides in association with lipids and proteins, represent the main constituent of
fungal cell wall. In filamentous fungi, outer cell wall layers mainly contain neutral
polysaccharides (glucans and mannans) while the inner layers contain more of
glucosamines (chitin and chitosan) in a microfibrillar structure. Ligands within these
matrices include carboxylate, amine phosphate, hydroxyl, Sulphydral and another
functional group. Proteins are also found to be associated with metal binding.
Rhizopus arrhizus has been used for accumulation of uranium and thorium a common anion
exchange resin used for the accumulation of uranium. The study of uranyl ion, lanthanum
and other divalent transition metal ion by Rhizopus arrhizus identified phosphate and
carboxyl group as binding groups. Studies on uranium uptake by Aspergillus niger and
Saccharomyces cerevisiae have indicated involvement of carboxylic group.
5.2Biosorption by Algae and Moss
Photoautotroph marine algae have bulk availability of their biomass from water bodies.
Sargassum natans and Ascophyllum nodosum in this group have shown very high
biosorptive capacities for various metals. Besides marine algae there are reports on binding
of heavy metal to green algae Chlorella and Cyanobacteria. Special polysaccharides
present in the algae cell wall contain potential metal ion binding sites. The number and
kind of binding sites depends on the chemical composition of the cell wall. In pheophycean
members, algin is present and contributes significantly to metal binding. It was suggested
that the polysaccharides of cell and could provide amino and carboxyl group as well as the
sulphate. The amino, carboxyl group and the nitrogen and oxygen-based moieties could

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also form coordinated and with metal ion. Metal ion could also be electrostatically bonded
to unprotected carboxyl oxygen and sulphate covalent bonding between divalent cation and
algae cell wall proteins has also been reported.
Mechanism such as entrapment of metal both in the form of insoluble microdeposits in the
inter and intra-fibrillar capillaries ad Para crystalline regions of polysaccharides and the
binging to biopolymers (RNA, polyphosphates) can contribute to the metal binding. The
groups are carboxylate, amine, imidazole, phosphate sulfhydryl, sulfhydryl, sulfate and
hydroxyl. Of these amides and imidazole’s are positively charged when protonated and
build negatively charged are also available for coordination bonding with metal ions such
as Lead (II) copper (II) and chromium (VI). Such bond formation could be accompanied
by displacement of protons and is dependent in part on extent of protonation, which is
determined by the pH.
5.3Biosorption by Bacteria
Bacteria may carry determinants of resistance to a number of heavy metals, including Ag,
Bi, Cd, Cu, Co, Hg, Pb, Ni, Zn cations and the oxyanions of As, Cr, Sb, such resistance
determinants have been extensively reviewed in recent years. Bacteria; resistance to heavy
metals in conferred by specific resistance determinants, which are often but not always
carried on plasmids or transposons. Resistance is specific to one or a few metals, and the
mechanisms of resistance include efflux of the metal, modification of the specification of
the metal sequestration of the metal or a combination of these mechanisms. The
mechanism of heavy metal resistance has indicated that similar mechanisms for resistance
to a single metal may occur across a wide range of bacterial genera and that related
mechanism of resistance may apply to different heavy metals. Living organisms have been
in intimate contact with heavy metals released into the environment by geochemical
process since organism first evolved, and over geological time scales microorganisms have
evolved to occupy ecological niches containing high concentration of heavy metals. Those
similar mechanisms being selected across different bacterial genera and for different metals
are not surprising. The proteins conferring metal resistance, the genetic regulatory

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mechanisms and bioremediation and biomonitoring strategies for environments which are
contaminated with heavy metals. Cell walls of bacteria and cyanobacteria are principally
composed of gram-negative bacteria is not heavily cross linked. They have an outer
membrane which is gram negative bacteria are more widespread in metal contaminated
soils than gram positive electrostatic interaction. Because of their thickness and anionic
character which is mainly due to high capacity for metal binding. It has been reported that
peptidoglycan in their major cell wall component responsible for meal binding by Bacillus
subtilis. In contrast Bacillus lichenifornnis metal binding is predominately due to
teichuronic acid. Among bacteria Bacillus species has been used for bio sorbent
preparation.
6.THE FACTORS AFFECT THE BIOSORPTION PROCESS
1.The temperature seems not to influence the biosorption performances in the range of
20-30°C
2. pH seems to be the most important parameter in the biosorptive process: it affects the
solution chemistry of the metals the activity of the functional groups in the biomass and
the competition of metallic ions
3. Biomass concentration in solution seems to influence the specific uptake: the
responsibility of the specific uptake decreases to metal concentration shortage in solution.
Hence this factor needs to be taken into consideration in any application of microbial
biomass as bio sorbent.
4.Biosorption is mainly used to treat waste water where more than one type of metal ions
would be present the removal of one metal ion may be influenced by the presence other
metal ion may be influenced by the presence of other metal ions. For example – Uranium

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uptake by biomass of bacteria, fungi and yeasts was not affected by the presence of
manganese, cobalt, copper, cadmium, mercury and led solution. In contrast the presence
of Fe2+ and Zn2+ was found in influence uranium uptake by
Rhizopus arrhizus and cobalt uptake by different microorganism seemed by completely
inhibited by the presence of uranium, lead, mercury, and copper.
6.1 Microorganisms used in Biosorption Process
Table -2: list of microorganisms used in Biosorption Process
Metals Micro Organisms employed
Arsenic Kocuria sp.
Bacillus sp.
Pseudomonas sp.
Cadmium Cupriavidus sp.
Enterobacter sp.
Stenotrophomonas sp.
Actinomycetes sp.
Chromium Staphylococcus sp.
Enterobacter sp.
Pseudomonas sp.
Micrococcus sp.
Cobalt Rhodopsudomonas sp.
Stenotryphomonas sp.

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Bacillus sp.
Gcobacillus sp.
Copper Bacillus sp.
Thiobacillus sp.
Enterobacter sp.
Lead Enterobacter sp.
Micrococcus sp.
Gcobacillus sp.
7. TOXITY OF HEAVY METALS – HUMANS
Most heavy metals are distributed in the body through blood to tissues. Lead is carried by red blood
cells to the liver and kidney and subsequently redistributed to the teeth, bone and hair mostly as
phosphate salt. Cadmium initially binds to blood cells and albumin and subsequently binds to
metallothionein in kidney and liver tissue. Following its distribution from blood to the lungs manganese
vapor diffuses across the lung membrane to the central nervous system. Organic salts of manganese
which are liquid soluble are distributed in the intestine for fecal elimination while inorganic manganese
salts which ae water soluble are distributed in plasma and kidney for renal elimination. Arsenic is
distributed in blood and accumulates in heart, lung, liver, kidney muscle and neural tissues and also in
the skin, nails and hair.
Living organisms require varying amounts of heavy metals. Iron, cobalt, copper, manganese,
molybdenum, and zinc are required by humans all metals are toxic at higher concentrations. Excessive
levels can be damaging the organism. Other heavy metals such as mercury, plutonium and lead are toxic
metals that have no known vital or beneficial effect on organisms, and their accumulation over time in
the bodies of animals can cause serious illness. Certain elements that are normally toxic are for certain
organism or under certain conditions beneficial. Example include vanadium, tungsten, and even
cadmium.

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Minamata Incident (1950)
Large amount of organic mercury was dumped into Minamata Bay. Mercury contaminated fish was
consumed by pregnant women newborn were with severe nerve damage. It was later known as Fetal
Minamata Disease.
Table -2: Types of heavy metals and their effect on human health with permissible limits
Pollutants Effect on human health Permissible level (mg/l)
Arsenic Bronchitis, Dermatitis, Poisoning 0.02
Cadmium Renal dysfunction, Lung disease, Lung
cancer, Bone defects, increased blood
pressure, kidney damage, bronchitis,
gastrointestinal disorder, bone marrow,
cancer
0.06
Lead Mental retardation in children,
developmental delay, fatal infant
encephalopathy, congenital paralysis,
sensor neural deafness and acute or
chronic damage to the nervous system,
epilepticus, liver, kidney,
gastrointestinal damage
0.1
Manganese Inhalation or contact causes damage to
central nervous system
0.26
Mercury Pesticides, batteries, paper industry 0.01
Zinc Refineries, brass manufacture, metal
plating, Plumbing
15
Chromium Mines, mineral sources 0.05
Copper Mining, Pesticide Production,
Chemical industry, metal piping
0.1

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8.CONCLUSION
Biosorption is eco-friendly and cheap method of removing metals form the environment. Previous
researches conducted during last five decades provided vast amount of information about different types
of biosorbents and their mechanism of metal uptake. More research is needed to explore new biosorbents
from environment. A deep insight is required not only on method of metal removal, but also its efficient
recovery so that it can be obtained into usable form.

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9.References:
1.Badriyah shaded Alotaibi, Maryam Khan, Saba Shamin; Unraveling the underlying
Heavy Metal Detoxification Mechanisms of Bacillus Species; 2021
2.Mounika Priyadarshanee, Surajit Das, Biosorption and removal of toxic of toxic heavy
metals by metal tolerating bacteria for bioremediation of metal contamination; 2020
3.Ayansina segun Ayanbenro and Olubukola Oluranti Babalola; A New Strategy for Heavy
Metal polluted Environments- A Review of Microbial Biosorbents; 2017
4.Salman H Abbas, Ibrahim M Ismail, Tarek M Mostafa, Abbas H. Sulaymon; Biosorption
of Heavy Metals; A Review
5.AnYan, Yamin Wang, Swee Ngin Tan, Mohamed Lokman Moh Yusof, Subhadip Ghosh
and Zhong Chen; Phytoremediation: A Promising Approach for Revegetation of Heavy
Metal Polluted Land; 2020
6.Introduction to Environmental Biotechnology, Third Edition; A.K.Chatterji: 163-165
7.Text Book of Environmental Biotechnology; Pradipta Kumar Mohapatra:412-437
8.Environmental Biotechnology Basic Concepts and Applications, Second Edition; Indu
Shekhar Thakur :312-318

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DUDEKULA HABEEBUNNISA